The highly divergent spermatozoon of the Palawan spiny rat, Maxomys panglima has an extremely long tail.

CONTEXT: Sperm morphology varies greatly across mammalian species and this variability is especially evident in murid rodents with both sperm head shape and tail length being sexually selected traits. The Palawan spiny rat, Maxomys panglima has a longer sperm tail than that currently recorded for any other mammalian species. AIMS: The aim of the current study was to determine the sperm morphology of an individual Palawan spiny rat, M. panglima . METHODS: Light and transmission electron microscopy were carried out. KEY RESULTS: We found that the sperm tail of M. panglima has an average length of 380µm with the midpiece being approximately 185µm in length with comparatively small mitochondria but very large coarse fibres. Furthermore, the sperm head has a less acutely flexed apical hook than that of most other murid rodents including those of several other Maxomys species. CONCLUSIONS: The Palawan spiny rat has a highly divergent sperm morphology with an extremely long tail. It may turn out to be an important species for testing various hypotheses of sperm form and function in mammals. IMPLICATIONS: These findings suggest markedly different selective pressures may have resulted in this unique sperm morphology, the functional significance of which remains to be determined.

Necropsy findings of koalas from the Mount Lofty Ranges population in South Australia.

OBJECTIVE: This study reports necropsy findings of koalas from the Mount Lofty Ranges region in order to identify health threats to this mainland South Australian population. METHODS: Koalas from the Mount Lofty Ranges region (n = 85) that had died or been euthanased on welfare grounds were examined at necropsy during 2012-13 at the School of Animal and Veterinary Sciences, University of Adelaide. Disease findings, approximate age, sex and body condition of koalas were recorded. Histopathological examination was undertaken on gross lesions and in suspect cases, skin scrapings taken for microscopy and PCR performed for Chlamydia pecorum detection. RESULTS: Traumatic injury was the most common necropsy finding (48/85; 57%), caused by motor vehicle accidents (35/48; 73%), canine attacks (11/48; 23%) or bushfire burns (2/48; 4%). Oxalate nephrosis (27/85; 32%) was also more common than other conditions. Infectious diseases included chlamydiosis (10/85; 12%) and sarcoptic mange (7/85; 8%). Marked testis asymmetry was evident in 11% (6/56) of males, with histopathology suggestive of atrophic change in four animals. Other pathological conditions included gastrointestinal disease (7/85; 8%) and respiratory disease (3/85; 4%). Almost half of the koalas (38/85; 45%) were found to have two or more abnormalities at necropsy. CONCLUSION: This study found trauma, mainly from motor vehicle accidents, and oxalate nephrosis to be the predominant causes of death and/or disease in koalas from the Mount Lofty Ranges region. Recent emergence of both clinical chlamydiosis and sarcoptic mange has also occurred, providing insight into the health status and causes of disease or injury in this South Australian mainland koala population.

Effects of whole-body heat on male germ cell development and sperm motility in the laboratory mouse.

This study investigated the effects of high temperatures on male germ cell development and epididymal sperm motility of laboratory mice. In Experiment 1, adult males (n=16) were exposed to whole-body heat of 37-38°C for 8h day(-1) for 3 consecutive days, whereas controls (n=4) were left at 23-24°C. In Experiment 2, adult mice (n=6) were exposed to 37-38°C for a single 8-h period with controls (n=6) left at 23-24°C. Experiment 2 was conducted as a continuation of previous study that showed changes in spermatozoa 16h after exposure to heat of 37-38°C for 8h day(-1) for 3 consecutive days. In the present study, in Experiment 1, high temperature reduced testes weights 16h and 14 days after exposure, whereas by Day 21 testes weights were similar to those in the control group (P=0.18). At 16h, 7 and 14 days after exposure, an increase in germ cell apoptosis was noticeable in early and late stages (I-VI and XI-XII) of the cycle of the seminiferous epithelium. However, apoptosis in intermediate stages (VII-X) was evident 16h after heat exposure (P<0.05), without any change at other time periods. By 21 days, there were no significant differences between heat-treated groups and controls. Considerably more caspase-3-positive germ cells occurred in heat-treated mice 16h after heat exposure compared with the control group (P<0.0001), whereas 8h after heat in Experiment 2, sperm motility was reduced with a higher percentage of spermatozoa showing membrane damage. In conclusion, the present study shows that whole-body heat of 37-38°C induces stage-specific germ cell apoptosis and membrane changes in spermatozoa; this may result in reduced fertility at particular times of exposure after heating.

Scalp cooling to prevent alopecia after chemotherapy can be considered safe in patients with breast cancer.

With modern scalp cooling equipment cytotoxic damage of hair root cells can be prevented in half of the patients with cancer at high risk of alopecia. However, traditionally doubt has existed whether scalp cooling might facilitate hiding and disseminating scalp skin metastases and thus decrease survival. We discuss this risk using frequency data on metastases in breast cancer from observational and autopsy studies and the Munich cancer registry. They showed the incidence of scalp skin metastases to be very low and not differ between scalp-cooled (0.04-1%) and non scalp-cooled (0.03-3%) patients with breast cancer and in need of chemotherapy. We found it rather unlikely that the incidence of scalp skin metastases might increase at all after scalp cooling, whereas a very small proportion of patients receiving chemotherapy are at risk to develop metastases at this site. Scalp cooling can thus safely be offered to patients treated with alopecia-inducing chemotherapy.

Impact of scalp cooling on chemotherapy-induced alopecia, wig use and hair growth of patients with cancer.

INTRODUCTION: Cytotoxic therapy for patients with cancer frequently induces reversible, but long-lasting alopecia which might be prevented by scalp cooling. This study evaluates the effectiveness of scalp cooling with respect to the severity of chemotherapy-induced alopecia (CIA) and the purchase and use of wigs and head covers. MATERIALS AND METHODS: In this observational study, scalp-cooled patients (n = 160) were compared with non scalp-cooled patients (n = 86) with several types of cancer. Patients were enrolled in 15, mostly general hospitals prior to taxane and/or anthracycline-based chemotherapy. Patients completed four questionnaires between the start and one year after the last chemotherapy. RESULTS: Severity of CIA, and purchasing and actually wearing wigs and head covers were significantly lower among scalp-cooled than non scalp-cooled patients. Overall, scalp cooling reduced the use of wigs and head covers by 40%. Among 84 scalp-cooled patients who purchased a wig (53%), only 52 patients actually wore it (62%), and they just wore it intensively (86% daily) for less than six months (80%). Especially young patients camouflaged CIA with a head cover instead of a wig. DISCUSSION: The relatively long duration of CIA, the wish of many patients to camouflage or rather prevent it and the 40% reduction for head covering by scalp cooling, makes it a worthwhile supportive intervention. However, (cost-) effectiveness can be improved. Many scalp-cooled patients purchased a wig unnecessarily.

Pathological features of oxalate nephrosis in a population of koalas (Phascolarctos cinereus) in South Australia.

The wild and captive koala population of the Mt Lofty Ranges in South Australia has a high level of renal dysfunction in which crystals consistent with calcium oxalate have been observed in the kidneys. This study aimed to describe the pathological features of the renal disease in this population, confirm the composition of renal crystals as calcium oxalate, and determine whether any age or sex predispositions exist for this disease. A total of 51 koalas (28 wild rescues, 23 captive) were examined at necropsy, of which 28 (55%) were found to have gross and/or histological evidence of oxalate nephrosis. Histopathological features included intratubular and interstitial inflammation, tubule dilation, glomerular atrophy, tubule loss, and cortical fibrosis. Calcium oxalate crystals were demonstrated using a combination of polarization microscopy, alizarin red S staining, infrared spectroscopy, and energy-dispersive X-ray analysis with scanning electron microscopy. Uric acid and phosphate deposits were also shown to be present but were associated with minimal histopathological changes. No significant differences were found between the numbers of affected captive and wild rescued koalas; also, there were no sex or age predispositions identified, but it was found that oxalate nephrosis may affect koalas <2 years of age. The findings of this study suggest that oxalate nephrosis is a leading disease in this koala population. Possible causes of this disease are currently under investigation.

Short post-infusion scalp cooling time in the prevention of docetaxel-induced alopecia.

PURPOSE: The patient impact of chemotherapy-induced alopecia (CIA) is high. Scalp cooling is applied to reduce CIA. The potential optimum post-infusion cooling times (PICTs) are currently unknown. METHODS: Scalp cooling was applied in 53 patients receiving docetaxel chemotherapy with 90-min PICT (observational part). Also 15 non-scalp-cooled patients were included. If hair preservation was observed in >80 % of the patients, randomisation between 45 and 90-min PICT was planned. Patients reported tolerance of scalp cooling and use of head covering. RESULTS: Observational study: 81 % of scalp-cooled patients did not require head covering versus 27 % of non-scalp-cooled patients. Randomised study: 79 % of 38 patients with 90-min PICT did not need head covering versus 95 % of 38 patients with 45-min PICT (p = 0.04). Scalp cooling was very well tolerated (visual analogue scale = 79). CONCLUSION: A 45-min PICT can be recommended in 3-weekly docetaxel regimens with a dose of 75 or 100 mg/m(2), administered in 60 min. The shorter PICT is a major advantage in time investment for patients. Patients (women and men) who receive docetaxel, except combined with doxorubicin and cyclophosphamide (taxotere, adriamycin and cyclophosphamide (TAC)) should be informed about the protective effect and high tolerability of scalp cooling in avoiding CIA.

Sperm morphology in the Malagasy rodents (Muroidea: Nesomyinae).

The morphology of the spermatozoon of representative species of the subfamily Nesomyinae (Muroidea: Nesomyidae), a monophyletic group of rodents endemic to Madagascar, was examined by light and electron microscopy to determine the sperm head shape and tail length across the species. Marked interspecific differences were found to occur in both the form of the sperm head and length of the tail. The species that possess a sperm head with an apical hook, which largely contains acrosomal material, generally displayed longer sperm tails, and a species with a spatulate sperm head had the shortest tail. The association between sperm head shape and tail length mirrors that previously found in Eurasian and Australasian murine rodents. Thus, the repeated association between sperm head shape and tail length across these groups of muroid rodents clearly indicates a functional relationship between these two features. A comparison of sperm morphology of the nesomyines to that of related muroid rodents on the mainland of Africa suggests that the possession of an apical hook is the ancestral condition.

The mechanistic basis for sexual dysfunction in male transforming growth factor beta1 null mutant mice.

The cytokine transforming growth factor beta1 (TGFB1) is implicated in male sexual function. Previous behavioral studies show that Tgfb1 null mutant mice mount and display limited intromission behavior with receptive females but are unable to complete successful copulation. The studies presented here explore the physiologic basis for sexual dysfunction in Tgfb1 null mutant males. Scanning electron microscopy revealed that the surface of the penis in Tgfb1 null mutant males was abnormally coated in superficial keratinized epithelial cells. There was a significant reduction in protrusion of penile spines through the superficial tissue in Tgfb1 null mutant mice; in some mice, the spines were almost completely embedded. Histologic analysis revealed reduced skin thickness in the penis of Tgfb1 null mutant males. Nerve fibers, endothelial cells, smooth muscle actin, macrophages, and neuronal and inducible nitric oxide synthase were present in similar abundance and location in Tgfb1 null mutant mice compared with wild-type controls; however, an increase in collagen I deposition was detected. Behavioral studies revealed that Tgfb1 null mutant males undergo spontaneous noncontact erections, albeit at a reduced rate compared with control mice, and engage in less frequent genital grooming activity. These studies suggest that Tgfb1 null mutation may adversely influence copulatory behavior through effects on both altered structural integrity of the penile skin and impaired tissue compliance leading to erectile dysfunction.

Spawning dynamics and biomass estimates of an anchovy Engraulis australis population in contrasting gulf and shelf environments.

The spawning biomass of Australian anchovy Engraulis australis in gulf and shelf waters of South Australia was compared using the daily egg production method (DEPM). The total survey area was 128 700 km2 with recorded spawning areas in gulf and shelf waters of 4898 and 44 618 km2, respectively. High egg densities in the warm, shallow gulf waters were produced by small, young (<1 year old) E. australis that spawned relatively small batches of eggs (c. 855) approximately every 3 days. In cooler, deeper shelf waters, where larger, older E. australis are found, lower egg densities occurred despite individuals producing much larger batches of eggs (c. 15,572) approximately every 7 days. In shelf waters, the highest densities were recorded at inshore sampling stations. Spawning appeared to peak between 0000 and 0100 hours. Females were more abundant than males in samples from both gulf and shelf waters with sex ratios of 0.61 and 0.56, respectively. The spawning biomass of E. australis in shelf waters was 101 522 t, whereas the estimate for gulf waters was 25 374 t. Due to the differences in mean size of the spawning females, however, c. 6x10(9)E. australis were present in each region. The results support the hypothesis that variability in habitat conditions may directly influence E. australis reproduction. A large reserve of young fish in the relatively stable gulf environment may increase the resilience of the E. australis population in South Australia to unfavourable interannual changes in offshore environmental conditions.

Ovarian follicular superstimulation and oocyte maturation in the anoestrous southern hairy-nosed wombat, Lasiorhinus latifrons.

This study investigates the effect of three exogenous gonadotrophin regimens on ovarian follicular development in southern hairy-nosed wombats during the non-breeding season. Females were given either porcine follicle stimulating hormone (pFSH; total of 200 mg at 12 h intervals over 7 (Group 1), or 4 days (Group 2)), or pregnant mares' serum gonadotrophin (PMSG; single dose of 150 I.U. (Group 3)). In all treatment groups 25 mg of porcine luteinising hormone (pLH) was used to trigger maturation; Groups 1 and 2 received pLH 12 h after the final pFSH injection and Group 3 received pLH 72 h after PMSG. The results showed Group 1 produced significantly more follicles per ovary (5.91+/-1.28) than Group 2 (1.67+/-0.62), or Group 3 (2.17+/-1.16) at p<0.05. Control females received saline injections concurrently with the three treatment groups (n=6; 2 control animals for each treatment group). No follicular development occurred in any control female. Analysis of oocyte nuclear status revealed that while oocytes from all three treatment groups had resumed meiosis, only those in Group 1 (7-day pFSH/pLH treatment) progressed to metaphase II. These results have implications for the development of assisted breeding strategies in this species.

Variation of sperm head shape and tail length in a species of Australian hydromyine rodent: the spinifex hopping mouse, Notomys alexis.

In Australia, there are around 60 species of murid rodents that occur in the subfamily Hydromyinae, most of which produce highly complex, monomorphic, spermatozoa in which the head has an apical hook together with two ventral processes containing filamentous actin and a long tail of species-specific length. One of the few exceptions to this is the spinifex hopping mouse, Notomys alexis, whose spermatozoa have previously been shown to have pleiomorphic heads. In this study, the structural organisation of the sperm head has been investigated in more detail and the variability in length of the midpiece and total length of the sperm tail has been determined for this species. The findings confirm that pleiomorphic sperm heads are invariably present in these animals and that this variability is associated with that of the nucleus, although nuclear vacuoles were not evident. The total length of the sperm tail, as well as that of the midpiece, was also highly variable both within, as well as between, individual animals. The reason(s) for this high degree of variability in sperm morphology is not known but it may relate to a relaxation of the genetic control of sperm form owing to depressed levels of inter-male sperm competition.

Effect of exogenous gonadotrophins on ovarian morphology and oocyte maturation in the southern hairy nosed wombat Lasiohinus latifrons during the breeding season.

The effect of the exogenous administration of porcine follicle-stimulating hormone (pFSH) and pregnant mare serum gonadotrophin (PMSG) on ovarian follicular development and oocyte maturation in the southern hairy nosed wombat Lasiorhinus latifrons was investigated. Three experimental groups were administered pFSH at various doses and for different treatment lengths, followed by 25 mg porcine luteinising hormone (pLH) 12 h after the last dose of pFSH. Another group was given PMSG followed 72 h later by 25 mg pLH. Animals were killed 24 h after pLH. The left ovary was fixed for histology and the morphology of the antral follicles was determined, whereas follicular oocytes in the right ovary were aspirated, fixed, stained with 42,62-diamidino-2-phenylindole, and viewed for nuclear maturation. There was no significant difference in the mean number of ovarian follicles >1 mm, or in the size class of follicles assessed between control and experimental groups. However, a trend was observed suggesting a possible increase in follicles >3.0 mm in experimental groups compared with control animals. In all females administered exogenous porcine gonadotrophins, but not controls, some of the mural granulosa cells of large tertiary antral follicles had markedly enlarged nuclei (approximately 14 microm in diameter). All oocytes from the control group remained at the germinal vesicle stage, whereas approximately 40% of oocytes retrieved from the pFSH groups and 82.4% retrieved from the PMSG-primed animals had undergone germinal vesicle break down, with a small number reaching meiosis II. The present study shows that exogenous administration of either pFSH or PMSG to hairy nosed wombats can induce follicular growth and oocyte maturation. Such findings could be useful in the development of reproductive technology in this species.

Comparative study of sperm chromatin condensation in the excurrent ducts of the laboratory mouse Mus musculus and spinifex hopping mouse Notomys alexis.

In most mammals, post-testicular sperm maturation is completed in the caput and corpus epididymides, with storage occurring in the cauda epididymides. However, in the spinifex hopping mouse, Notomys alexis, epididymal sperm transit is rapid and some sperm storage occurs in the distal region of the vas deferens. The aim of the present study was to determine whether the rapid progression of sperm into the vas deferens in the hopping mouse results in late sperm maturation. To determine this, sperm nuclei from the epididymides and vasa deferentia of laboratory and hopping mice were compared for: (1) thiol content after staining with monobromobimane (mBBr); (2) chromatin resistance to acid denaturation following incubation with acetic alcohol and staining with acridine orange; and (3) chromatin resistance to in vitro decondensation after incubation with 1% sodium dodecyl sulfate (SDS). It was found that, whereas laboratory mouse sperm completed chromatin condensation by the time they reached the cauda epididymidis, hopping mouse sperm nuclei from the vas deferens showed significantly less mBBr fluorescence and a greater proportion of sperm were resistant to decondensation with SDS than those in the cauda epididymidis. Therefore, the results of the present study indicate that, unlike in the laboratory mouse, hopping mouse chromatin condensation of spermatozoa continues in the vas deferens and this may be due, at least in part, to rapid epididymal transit.

Morphogenesis of the fibrous sheath in the marsupial spermatozoon.

The spermatozoon fibrous sheath contains longitudinal columns and circumferential ribs. It surrounds the axoneme of the principal piece of the mammalian sperm tail, and may be important in sperm stability and motility. Here we describe its assembly during spermiogenesis in a marsupial, the brush-tail possum, and compare its structural organization with that of eutherian mammals, birds and reptiles. Transmission electron microscopy showed that possum fibrous sheath assembly is a multistep process extending in a distal-to-proximal direction along the axoneme from steps 4 to 14 of spermiogenesis. For the most part, assembly of the longitudinal columns occurs before that of the circumferential ribs. Immunohistochemical and immunogold labelling showed that fibrous sheath proteins are first present in the spermatid cytoplasm; at least some of the proteins of the sheath precursors differ from those in the mature fibrous sheath. That immunoreactivity develops after initiation of chromatin condensation suggests that fibrous sheath proteins, or their mRNAs, are stored within the spermatid cytoplasmic lobule prior to their assembly along the axoneme. These findings are similar to those in laboratory rats, and thus suggests that the mode of fibrous sheath assembly evolved in a common ancestor over 125 million years ago, prior to the divergence of marsupial and eutherian lineages.

Unusual germ cell organization in the seminiferous epithelium of a murid rodent from southern Asia, the greater bandicoot rat, Bandicota Indica.

In the greater bandicoot rat, Bandicota indica, of south-east Asia, nine cell associations were documented in the testicular seminiferous epithelium. In about 10% of the tubule cross sections two or more cell associations occurred and, furthermore, some of the generations of germ cells within the cell associations were sometimes either out of phase, or missing, in the tubule cross sections. These features, together with the fact that this species has a highly pleiomorphic sperm head shape, are somewhat reminiscent of those of the seminiferous epithelium in humans and some other primates but not of common laboratory rodents. This species could thus be a good model for investigating irregular patterns of spermatogenesis in naturally occurring wild species of rodent.

Interferon-alpha as maintenance therapy in patients with multiple myeloma.

BACKGROUND: The effect of interferon-alpha 2b (IFN-alpha-2b) on progression-free and overall survival as well as quality of life (QoL) was studied in mainly elderly patients with multiple myeloma (MM), who reached a plateau phase after melphalan/prednisone induction. PATIENTS AND METHODS: In an open phase III trial, 262 patients, median age 69 years (range 34-91), received at least 10 monthly courses of melphalan/prednisone followed by response evaluation. Plateau phase was reached by 128 patients. Next, 90 patients were randomized between IFN-alpha-2b and no maintenance therapy. Reasons for non-randomization were: refusal (18), concomitant disease (nine), protocol violation (six), WHO performance status >2 (four) and allogeneic transplantation (one) RESULTS: At a median follow-up from diagnosis of 97 months (0-140) for those patients alive, IFN-alpha-2b therapy was associated with improved progression-free survival (median 13.5 versus 8.4 months from randomization), although this did not translate in a better overall survival (41 versus 38.4 months). One-third of patients discontinued IFN-alpha due to toxicity. No differences were observed between patient groups in QoL. CONCLUSIONS: IFN maintenance therapy in MM prolongs progression-free survival and, provided that the burden of toxicity is not too high, does not adversely affect QoL.

Prevention of chemotherapy-induced hair loss by scalp cooling.

BACKGROUND: Chemotherapy-induced temporary hair loss is one of the most common and distressing side-effects of cancer therapy. Scalp cooling to reduce this hair loss is a controversial issue for many doctors and nurses. This may be due to inadequate knowledge. METHODS: This review from 53 publications and three personal communications focuses on the efficacy of the treatment, side-effects, possible disadvantages and the controversies in these areas. RESULTS: Scalp cooling has become an increasingly effective method to prevent hair loss, especially when anthracyclines or taxanes are used. Unfortunately, many studies were small and badly designed and are therefore difficult to compare. There is a considerable variation in the success rates in the various studies. This remains unexplained, but the cooling time, the chemotherapy used and the temperature seem to be influential. Scalp cooling should not be used if chemotherapy is given with a curative intent in patients with generalised haematogenic metastases. The majority of patients tolerate cooling very well. CONCLUSION: Scalp cooling is effective but not for all chemotherapy patients. Further psychological, clinical and biophysical research is needed to determine exact indications for cooling and to improve the effect, tolerance, side-effects and the cooling procedure. Multicentre trials should be carried out to gather this information.

Studies on sperm storage in the vas deferens of the spinifex hopping mouse (Notomys alexis).

The cauda epididymidis, with its relatively cool temperature (32-35 degrees C), is considered to be the main site of sperm storage in male mammals. However, in the adult male spinifex hopping mouse, Notomys alexis, similar numbers of spermatozoa are found in the vas deferens to those in the cauda epididymidis. The present study shows that, unlike in the laboratory mouse in which spermatozoa of the vas deferens are found mainly in the epididymal region of the duct, spermatozoa in the hopping mouse are localized mainly to the middle and urethral regions of the vas deferens which lies in the inguinal and lower abdominal region of the body cavity. After ligation of the vas deferens close to its connection with the epididymis, many spermatozoa in the vas deferens retain the potential for motility for up to 2 weeks, indicating that the viability of spermatozoa is not compromised by being restricted to core body temperature. This urethral region of the vas deferens, in which spermatozoa reside, has a highly divergent structural organization compared with that of common laboratory rodents in which there is an expanded lumen with a network of epithelial folds. Ultrastructural observations of the cells lining the duct indicate that there are not any marked differences in morphology compared with the cells lining the duct in common laboratory murids, but the infoldings of the vas deferens of the hopping mouse are highly vascular which might facilitate supply of oxygen and nutrients to the spermatozoa residing in the lumen.